Disorders of iron metabolism are among the most common human diseases. During the last decade, rapid progress has been made in the understanding of the molecular basis or iron homeostasis and its disorders. The peptide hormone hepcidin has emerged as the master regulator of iron metabolism. Dysregulation of hepcidin is the principal or contributing factor in most systemic iron disorders. There is a well recognized unmet need for a robust and widely available clinical assay for serum hepcidin but progress towards this goal has been slowed by the poor immunogenicity of hepcidin. Intrinsic Life Sciences (ILS) developed and validated the world's first serum hepcidin immunoassay (competitive ELISA), that relied on a finite supply of high quality rabbit polyclonal antibodies. In order to increase the availability and standardization of the hepcidin ELISA, Phase I SBIR funding was used to produce murine monoclonal antibodies against hepcidin. Under Phase II of this proposal we will: 1) Use the monoclonal antibodies to optimize and validate a 2nd generation improved competitive ELISA for hepcidin 2) Produce additional monoclonal antibodies to hepcidin and develop and validate a two-monoclonal antibody sandwich ELISA. Successful completion of Phase II will enable ILS to deliver a hepcidin immunoassay prototype suitable for FDA approval and widespread adoption by clinicians and researchers. PUBLIC HEALTH RELEVANCE: With Phase II SBIR funding, Intrinsic Life Sciences (ILS) will develop the first prototype diagnostic tests suitable for FDA-approval for the iron-regulating hormone hepcidin. Measurement of hepcidin will permit greatly improved diagnosis of prevalent genetic and acquired iron disorders and anemias associated with inflammatory diseases and cancers.